Detection of Escherichia coli O157:H7 by multiplex PCR and their characterization by plasmid profiling, antimicrobial resistance, RAPD and PFGE analyses

Twenty-five and three strains of Escherichia coli O157:H7 were identified from 25 tenderloin beef and three chicken meat burger samples, respectively. The bacteria were recovered using the immunomagnetic separation procedure followed by selective plating on sorbitol MacConkey agar and were identifie...

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Main Authors: Radu, Son, Ooi, Wai Ling, Rahmat Ali, Gulam Rusul, Abdul Karim, Mohamed Ismail, Nishibuchi, Mitsuaki
Format: Article
Language:English
English
Published: Elsevier 2001
Online Access:http://psasir.upm.edu.my/id/eprint/15296/1/Detection%20of%20Escherichia%20coli%20O157H7%20by%20multiplex%20PCR%20and%20their%20characterization%20by%20plasmid%20profiling%2C%20antimicrobial%20resistance%2C%20RAPD%20and%20PFGE%20analyses.pdf
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spelling my.upm.eprints.152962024-07-26T07:48:39Z http://psasir.upm.edu.my/id/eprint/15296/ Detection of Escherichia coli O157:H7 by multiplex PCR and their characterization by plasmid profiling, antimicrobial resistance, RAPD and PFGE analyses Radu, Son Ooi, Wai Ling Rahmat Ali, Gulam Rusul Abdul Karim, Mohamed Ismail Nishibuchi, Mitsuaki Twenty-five and three strains of Escherichia coli O157:H7 were identified from 25 tenderloin beef and three chicken meat burger samples, respectively. The bacteria were recovered using the immunomagnetic separation procedure followed by selective plating on sorbitol MacConkey agar and were identified as E. coli serotype O157:H7 with three primer pairs that amplified fragments of the SLT-I, SLT-II and H7 genes in PCR assays. Susceptibility testing to 14 antibiotics showed that all were resistant to two or more antibiotics tested. Although all 28 strains contained plasmid, there was very little variation in the plasmid sizes observed. The most common plasmid of 60 MDa was detected in all strains. We used DNA fingerprinting by randomly amplified polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) to compare the 28 E. coli O157:H7 strains. At a similarity level of 90%, the results of PFGE after restriction with XbaI separated the E. coli O157:H7 strains into 28 single isolates, whereas RAPD using a single 10-mer oligonucleotides separated the E. coli O157:H7 strains into two clusters and 22 single isolates. These typing methods should aid in the epidemiological clarification of the E. coli O157:H7 in the study area. Elsevier 2001 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/15296/1/Detection%20of%20Escherichia%20coli%20O157H7%20by%20multiplex%20PCR%20and%20their%20characterization%20by%20plasmid%20profiling%2C%20antimicrobial%20resistance%2C%20RAPD%20and%20PFGE%20analyses.pdf text en http://psasir.upm.edu.my/id/eprint/15296/7/1-s2.0-S016770120100269X-main.pdf Radu, Son and Ooi, Wai Ling and Rahmat Ali, Gulam Rusul and Abdul Karim, Mohamed Ismail and Nishibuchi, Mitsuaki (2001) Detection of Escherichia coli O157:H7 by multiplex PCR and their characterization by plasmid profiling, antimicrobial resistance, RAPD and PFGE analyses. Journal of Microbiological Methods, 46 (2). pp. 131-139. ISSN 0167-7012; ESSN: 1872-8359 https://www.sciencedirect.com/science/article/pii/S016770120100269X?via%3Dihub#! 10.1016/S0167-7012(01)00269-X
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
English
description Twenty-five and three strains of Escherichia coli O157:H7 were identified from 25 tenderloin beef and three chicken meat burger samples, respectively. The bacteria were recovered using the immunomagnetic separation procedure followed by selective plating on sorbitol MacConkey agar and were identified as E. coli serotype O157:H7 with three primer pairs that amplified fragments of the SLT-I, SLT-II and H7 genes in PCR assays. Susceptibility testing to 14 antibiotics showed that all were resistant to two or more antibiotics tested. Although all 28 strains contained plasmid, there was very little variation in the plasmid sizes observed. The most common plasmid of 60 MDa was detected in all strains. We used DNA fingerprinting by randomly amplified polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) to compare the 28 E. coli O157:H7 strains. At a similarity level of 90%, the results of PFGE after restriction with XbaI separated the E. coli O157:H7 strains into 28 single isolates, whereas RAPD using a single 10-mer oligonucleotides separated the E. coli O157:H7 strains into two clusters and 22 single isolates. These typing methods should aid in the epidemiological clarification of the E. coli O157:H7 in the study area.
format Article
author Radu, Son
Ooi, Wai Ling
Rahmat Ali, Gulam Rusul
Abdul Karim, Mohamed Ismail
Nishibuchi, Mitsuaki
spellingShingle Radu, Son
Ooi, Wai Ling
Rahmat Ali, Gulam Rusul
Abdul Karim, Mohamed Ismail
Nishibuchi, Mitsuaki
Detection of Escherichia coli O157:H7 by multiplex PCR and their characterization by plasmid profiling, antimicrobial resistance, RAPD and PFGE analyses
author_facet Radu, Son
Ooi, Wai Ling
Rahmat Ali, Gulam Rusul
Abdul Karim, Mohamed Ismail
Nishibuchi, Mitsuaki
author_sort Radu, Son
title Detection of Escherichia coli O157:H7 by multiplex PCR and their characterization by plasmid profiling, antimicrobial resistance, RAPD and PFGE analyses
title_short Detection of Escherichia coli O157:H7 by multiplex PCR and their characterization by plasmid profiling, antimicrobial resistance, RAPD and PFGE analyses
title_full Detection of Escherichia coli O157:H7 by multiplex PCR and their characterization by plasmid profiling, antimicrobial resistance, RAPD and PFGE analyses
title_fullStr Detection of Escherichia coli O157:H7 by multiplex PCR and their characterization by plasmid profiling, antimicrobial resistance, RAPD and PFGE analyses
title_full_unstemmed Detection of Escherichia coli O157:H7 by multiplex PCR and their characterization by plasmid profiling, antimicrobial resistance, RAPD and PFGE analyses
title_sort detection of escherichia coli o157:h7 by multiplex pcr and their characterization by plasmid profiling, antimicrobial resistance, rapd and pfge analyses
publisher Elsevier
publishDate 2001
url http://psasir.upm.edu.my/id/eprint/15296/1/Detection%20of%20Escherichia%20coli%20O157H7%20by%20multiplex%20PCR%20and%20their%20characterization%20by%20plasmid%20profiling%2C%20antimicrobial%20resistance%2C%20RAPD%20and%20PFGE%20analyses.pdf
http://psasir.upm.edu.my/id/eprint/15296/7/1-s2.0-S016770120100269X-main.pdf
http://psasir.upm.edu.my/id/eprint/15296/
https://www.sciencedirect.com/science/article/pii/S016770120100269X?via%3Dihub#!
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